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Genomic modelling of the ESR1 Y537S mutation for evaluating function and new therapeutic approaches for metastatic breast cancer

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Title: Genomic modelling of the ESR1 Y537S mutation for evaluating function and new therapeutic approaches for metastatic breast cancer
Authors: Harrod, A
Fulton, J
Nguyen, VTM
Periyasamy, M
Ramos Garcia, L
Lai, C-F
Metodieva, G
De Giorgio, A
Williams, RL
Santos, DB
Jimenez Gomez, P
Lin, M-L
Metodiev, MV
Stebbing, J
Castellano, L
Magnani, L
Coombes, RC
Buluwela, L
Ali, S
Item Type: Journal Article
Abstract: Drugs that inhibit estrogen receptor-α (ER) activity have been highly successful in treating and reducing breast cancer progression in ER-positive disease. However, resistance to these therapies presents a major clinical problem. Recent genetic studies have shown that mutations in the ER gene are found in >20% of tumours that progress on endocrine therapies. Remarkably, the great majority of these mutations localise to just a few amino acids within or near the critical helix 12 region of the ER hormone binding domain, where they are likely to be single allele mutations. Understanding how these mutations impact on ER function is a prerequiste for identifying methods to treat breast cancer patients featuring such mutations. Towards this end, we used CRISPR-Cas9 genome editing to make a single allele knockin of the most commonly mutated amino acid residue, tyrosine 537, in the estrogen-responsive MCF7 breast cancer cell line. Genomic analyses using RNA-seq and ER ChIP-seq demonstrated that the Y537S mutation promotes constitutive ER activity globally, resulting in estrogen-independent growth. MCF7-Y537S cells were resistant to the anti-estrogen tamoxifen and fulvestrant. Further, we show that the basal transcription factor TFIIH is constitutively recruited by ER-Y537S, resulting in ligand-independent phosphorylation of Serine 118 (Ser118) by the TFIIH kinase, CDK7. The CDK7 inhibitor, THZ1 prevented Ser118 phosphorylation and inhibited growth of MCF7-Y537S cells. These studies confirm the functional importance of ER mutations in endocrine resistance, demonstrate the utility of knockin mutational models for investigating alternative therapeutic approaches and highlight CDK7 inhibition as a potential therapy for endocrine resistant breast cancer mediated by ER mutations.
Issue Date: 17-Oct-2016
Date of Acceptance: 29-Aug-2016
URI: http://hdl.handle.net/10044/1/39675
DOI: https://dx.doi.org/10.1038/onc.2016.382
ISSN: 1476-5594
Publisher: Nature Publishing Group
Start Page: 2286
End Page: 2296
Journal / Book Title: Oncogene
Volume: 36
Copyright Statement: © The Author(s) 2016. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/ 4.0/ © The Author(s) 2016
Sponsor/Funder: Cancer Research UK
Funder's Grant Number: 14549
Keywords: Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Cell Biology
Genetics & Heredity
Antineoplastic Agents
Breast Neoplasms
CRISPR-Cas Systems
Cell Proliferation
Cyclin-Dependent Kinases
Estrogen Antagonists
Estrogen Receptor alpha
Gene Knock-In Techniques
MCF-7 Cells
1112 Oncology And Carcinogenesis
1103 Clinical Sciences
Oncology & Carcinogenesis
Publication Status: Published
Appears in Collections:Department of Surgery and Cancer